Biological significance of N-acetylglucosamine 6-sulfation

N-乙酰氨基葡萄糖6-硫酸化的生物学意义

• 批准号: 12480187
• 负责人: MURAMATSU Takashi
• 金额: $ 9.22万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12480187/
• 关键词: sulfotransferase; N-acetylglucosamine; L-selectin; Blood coagulation; Oligosaccharide synchesis; Glycoproteins; L-セレクチンリガンド; スルフォトランスフェラーゼ; リンパ球ホーミング; 癌性糖鎖; セレクチン; ルウィスX; 糖鎖抗原; 糖鎖生物学; 糖鎖識別

Three species of N-acetylglucosamine-6-sulfotransferases (GlcNAc6ST), namely GlcNAc6ST-1, GlcNAc6ST-2, GlcNAc6ST-3 were expressed and the specificities were compared. GlcNAc6ST-l as well as GlcNAc6ST-2 acted on mucin-type core 2 sugar (GlcNAcβ1-6[Galβ1-3]GalNAc) and there were not significant differences between the two enzymes. Although GlcNAc6ST-2 is known to be involved in lymphocyte homing, it is not clarified whether GlcNAc6ST-1 plays such a role. Cells doubly transfected with GlcNAc6ST-1 and fucosyltransferase VII induced roling of L-selectin-expressing cells. GlcNAc6ST-3 acted only on core 2 sugars. On the other hand, only GlcNAc6ST-2 acted on core 3 sugars (GlcNAcβ1-3 GalNAc). From this specificity, GlcNAc6ST-2 was considered to be the enzyme, which became newly expressed in mucinous adenocarcinoma, and this point was confined by RT-PCR analysis.We produced knockout mice deficient in the GlcNAc6ST-1 gene. That this mice completely lacked GlcNAc6ST-1 was confirmed by Southern and Northern blot analyses. The mice was born normally, and showed no apparent abnormality. Currently backcross is on the way, so that we are able to analyze possible abnormality in lymphocyte homing.GilcNAc6ST-1 was produced as fasion protein in Tn5 cells using the Baculovirus system. The purified recombinant enzyme was useful for synthesis of sulfated oligosaccharides and sulfation of glycoprotein-bound carbohydrates.

表达了三种N-乙酰葡糖胺-6-磺基转移酶(GlcNAc6ST)，即GlcNAc6ST-1、GlcNAc6ST-2、GlcNAc6ST-3，并比较了GlcNAc6ST-1和GlcNAc6ST-2作用于粘蛋白型核心2的特异性。糖(GlcNAcβ1-6[Galβ1-3]GalNAc)，两种酶之间没有显着差异，尽管已知 GlcNAc6ST-2 参与淋巴细胞归巢，但尚不清楚 GlcNAc6ST-1 是否发挥双重作用。用 GlcNAc6ST-1 和岩藻糖基转移酶 VII 转染诱导 L-选择素表达细胞的滚动。 GlcNAc6ST-3仅作用于核心2糖，而只有GlcNAc6ST-2作用于核心3糖(GlcNAcβ1-3 GalNAc)，因此认为GlcNAc6ST-2是新表达的酶。粘液腺癌，这一点受到 RT-PCR 分析的限制。我们制备了 GlcNAc6ST-1 基因缺陷的敲除小鼠。通过Southern和Northern印迹分析证实完全缺乏GlcNAc6ST-1的小鼠出生正常，并且没有表现出明显的异常，因此我们能够分析淋巴细胞归巢可能的异常。使用杆状病毒系统在 Tn5 细胞中作为 fasion 蛋白产生，纯化的重组酶可用于硫酸化寡糖的合成和硫酸化。糖蛋白结合碳水化合物。

项目成果

Chen G et al.: "A novel variant form of murine β-1,6-N-acetylglucosaminyl transferase forming branches in poly-N-acetyllactosamines."Glycobiology.. 10. 1001-1011 (2000)

Chen G 等人：“鼠 β-1,6-N-乙酰葡糖胺基转移酶的新型变体形式在聚 N-乙酰基乳糖胺中形成分支。”糖生物学.. 10. 1001-1011 (2000)

Kazuhiro Ishiguro: "Syndecan-4-Deficiency Leads to High Mortality of Lipopolysaccharide-injected Mice"The Journal of Biological Chemistry. Vol.276 No.50. 47483-47488 (2001)

Kazuhiro Ishiguro：“Syndecan-4-缺陷导致脂多糖注射小鼠的高死亡率”生物化学杂志。

Kenji Uchimura: "Specificities of N-acetylglucosamine-6-O-sulfotransferases in relation to L-selectin ligand synthesis and tumor-associated enzyme expression"The Journal of Biological Chemistry. 277. 3979-3984 (2002)

Kenji Uchimura：“N-乙酰葡糖胺-6-O-磺基转移酶与 L-选择素配体合成和肿瘤相关酶表达相关的特性”《生物化学杂志》。

Fathy M.El-Fasakhany: "N-Acetylglucosamine-6-sulfotransferase-1 : production in the baculovirus system and its applications to the synthesis of a sulfated oligosaccharide and to the modification of oligosaccharides in fibrinogen"Journal of Biochemistry. (

Fathy M.El-Fasakhany：“N-乙酰葡糖胺-6-磺基转移酶-1：杆状病毒系统中的生产及其在硫酸化寡糖合成和纤维蛋白原中寡糖修饰中的应用”生物化学杂志。

Kenji Uchimura et al.: "Specificities of N-acetylglucosamine-6-O-sulfotransferases in relation to L-selectin ligand synthesis and tumor-associated enzyme expression"The Journal of Biological Chemistry. 277. 3979-3984 (2002)

Kenji Uchimura 等人：“N-乙酰葡糖胺-6-O-磺基转移酶与 L-选择素配体合成和肿瘤相关酶表达相关的特异性”《生物化学杂志》。